| Anti-chlamydial antibodies and uses thereof -> Monitor Keywords |
|
|
  Anti-chlamydial antibodies and uses thereofRelated Patent Categories: Chemistry: Molecular Biology And Microbiology, Measuring Or Testing Process Involving Enzymes Or Micro-organisms; Composition Or Test Strip Therefore; Processes Of Forming Such Composition Or Test Strip, Involving Antigen-antibody Binding, Specific Binding Protein Assay Or Specific Ligand-receptor Binding Assay, Involving A Micro-organism Or Cell Membrane Bound Antigen Or Cell Membrane Bound Receptor Or Cell Membrane Bound Antibody Or Microbial Lysate, Bacteria Or ActinomycetalesThe Patent Description & Claims data below is from USPTO Patent Application 20060073531. Brief Patent Description - Full Patent Description - Patent Application Claims
CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation-in-part of U.S. patent application Ser. No. 10/020,269, filed Dec. 14, 2001, which is a continuation of U.S. patent application Ser. No. 09/025,596, filed Feb. 18, 1998, which is a continuation-in-part of U.S. patent application Ser. No. 08/911,593, filed Aug. 14, 1997. This application is also a continuation-in-part of U.S. patent application Ser. No. 10/873,768, filed Jun. 22, 2004, which is a continuation-in-part of U.S. patent application Ser. No. 08/911,593. U.S. patent application Ser. No. 08/911,593 claims benefit from U.S. provisional application U.S. Pat. Ser. No. 60/023,921, filed Aug. 14, 1996. The entire teachings of each of the foregoing applications are incorporated herein by reference. BACKGROUND OF THE INVENTION [0002] Antibodies are widely used in diagnostic assays in both human and veterinary medicine. Uses include enzyme-linked immunosorbent analysis (ELISA), quantitative antigen capture analysis, radioisotope-tagged reagents for in vivo localization of target antigens, and for in vivo localization of cytotoxic agents to target cells (i.e., immunotoxic therapy). The minimum epitope size for protein antigens is generally considered to be 5-6 amino acids, either as a linear sequence or as non-contiguous amino acids whose spatial placement defines the epitope (i.e., conformational epitope). Specificity is provided by the large number of potential amino acid epitopic sequences possible for a minimum epitope (i.e., 5.sup.20). [0003] Most commonly, large antigens or microbial organisms are used to induce antibody responses in order to insure the presentation of good antigenic sequences in the host animal. The use of these multivalent antigens for the production of polyclonal antibodies generally requires host-based adsorption of the sera to reduce non-specific cross-reactive antibody species. Monoclonal antibodies avoid this pitfall but frequently result in reagents whose specific epitopic specificity is unknown. SUMMARY OF THE INVENTION [0004] The invention relates to a method of identifying an antigenic amino acid subsequence from within a larger amino acid sequence comprising the steps of evaluating the hydrophilicity of subsequences of an amino acid sequence of interest; evaluating the flexibility of subsequences of the amino acid sequence of interest; and selecting an amino acid subsequence having overlapping regions of hydrophilicity and flexibility. In particular embodiments, the larger amino acid sequence is selected from the group consisting of polypeptides expressed by members of the Chlamydia genus. [0005] The invention also relates to antigenic amino acid subsequences identified by the methods described herein. In particular embodiments, the invention pertains to an antigenic amino acid subsequence selected from the group consisting of SEQ ID NOS: 1-118. [0006] The invention also relates to antibodies which are specific for the antigenic amino acid subsequences described herein. For example, the invention pertains to monoclonal antibodies specific for antigenic amino acid subsequences described herein. [0007] The invention also relates to diagnostic and therapeutic methods utilizing the described antigenic amino acid subsequences and antibodies thereto. [0008] The invention also encompasses a method for evaluating the infection status of an individual and/or the progress of therapy in an individual undergoing therapy for infection caused by Chlamydia. The method comprises quantifying antibody titer or other measure to the pathogen and comparing the measure to antibody measure quantified at a time earlier in the therapy, whereby the difference between the measures is indicative of the progress of the therapy. The invention also pertains to a method for monitoring the course of therapy for treating infection by Chlamydia, comprising determining presence or absence of Chlamydia in an infected individual at time intervals during course of therapy. In a particular embodiment, this is determined by PCR assay or antigen capture assay for pathogen DNA. [0009] Detection of the presence of Chlamydia in a sample of biological material taken from an individual thought to be infected therewith is important in determining the course of therapy and the agents to be used. This can be achieved by detecting the presence of DNA encoding MOMP of Chlamydia or other chlamydial genes in the individual. In one aspect of the invention, diseases associated with Chlamydia infection, such as inflammatory diseases, autoimmune diseases and diseases in which the individual is immunocompromised, can be treated by managing (e.g., significantly reducing infection or eradicating) the Chlamydia infection using the novel approach described herein. Both clinical and serological improvements/resolutions in patient status have been demonstrated. BRIEF DESCRIPTION OF THE DRAWINGS [0010] FIGS. 1A and 1B are sequence alignments of various Chlamydia MOMPs. Variable domains (VD1-VD4) are boxed. Sequences are aligned with the L2 serovar of C. trachomatis and are ranked from highest homology (B, D, E, L1) to lower homology (F, C, and A, H, L3). MU is the mouse pneumonitis C. trachomatis. PN refers to the human C. pneumonia. Deletions are indicated by (-). A blank indicates the same residue as L2. The leader sequence is bracketed. Underlined seven residue segments are predicted to contain the most flexible peptide backbone based on the L2 sequence. Asterisks indicate the most hydrophilic region. [0011] FIG. 2 illustrates the predicted antigenic sequences from variable domains 1 (VD1) of various Chlamydia species. The boxed cysteine (C) residue is not part of the native sequence but has been added at the amino terminus for cross-linking to carrier proteins used in immunization. [0012] FIG. 3 illustrates the predicted antigenic sequences from variable domain 2 (VD2) of various Chlamydia species. The boxed cysteine (C) residue is not part of the native sequence but has been added at the amino terminus for cross-linking to carrier proteins used in immunization. [0013] FIG. 4 illustrates the predicted antigenic sequences from a common domain of various Chlamydia species. The shaded box indicates hydrophilic mobile region common to each with expected cross-reactivity for antibodies specific for the sequence. The boxed cysteine (C) residue is not part of the native sequence but has been added at the amino terminus for cross-linking to carrier proteins used in immunization. DETAILED DESCRIPTION OF THE INVENTION [0014] Globular proteins have a hydrophobic core, with the external surfaces bearing relatively hydrophilic sequences. It is these segments in native proteins which are most likely to be recognized by antibodies. Work described herein describes methods for identifying linear amino acid antigenic sequences for the production of both polyclonal and monoclonal antibodies to defined antigenic domains. One significant advantage of this technique is that it provides antibodies to a known epitope of a target antigen or organism. [0015] The identification of antigenic domains described herein is based on the overlap of the most hydrophilic peptide segments of an antigen with those peptide segments with a concomitant predicted peptide flexibility. Increased flexibility allows more conformational degrees of freedom for optimal fit into an antibody binding site. Aromatic amino acids are frequently found in antigenic epitopes although hydrophobic with bulky R groups. This decrease in the relative hydrophobicity and flexibility of the peptide sequence containing the aromatic residue is compensated for if accessible (i.e., surface of the antigen). [0016] The relative hydrophilicity of peptide domains is based on the individual hydrophilicity of each amino acid in six or more residue segments as defined by Hopp and Woods (Hopp and Woods, Proceedings of National Academy of Sciences USA 78:3824-3828). Flexibility of the peptide chain at each C.alpha. residue is measured from the average value of the atomic temperature factor as affected by adjacent residues. Amino acids which result in rigidity of the chain include alanine, valine, leucine, isoleucine, tyrosine, phenylalanine, tryptophan, cysteine, methionine, and histidine. Flexibility is computed by averaging the rigidity factor (B value) along a seven residue segment using the following expression (Karplus and Shulz, Naturwissenschaften 72:212-213 (1985); Van Regenmortel, Trends in Biochemical Sciences (TIBS) 12:36-39 (1986)):F=B.sub.i+0.75(B.sub.i-1+B.sub.i+1)+0.5(B.sub.i-2+B.sub.i+2)+0.25- (B.sub.i-3+B.sub.i+3) [0017] With respect to identification of larger proteins or polypeptides from which the antigenic amino acid subsequences are selected, bands identified by gel analysis can be isolated and purified by HPLC, and the resulting purified protein can be sequenced. Alternatively, the purified protein can be enzymatically digested by methods known in the art to produce polypeptide fragments which can be sequenced. The sequencing can be performed, for example, by the methods of Wilm et al. (Nature 379:466-469 (1996)). The protein can be isolated by conventional means of protein biochemistry and purification to obtain a substantially pure product, i.e., 80, 95 or 99% free of cell component contaminants, as described in Jacoby, Methods in Enzymology Volume 104, Academic Press, New York (1984); Scopes, Protein Purification, Principles and Practice, 2nd Edition, Springer-Verlag, New York (1987); and Deutscher (ed), Guide to Protein Purification, Methods in Enzymology, Vol. 182 (1990). If the protein is secreted, it can be isolated from the supernatant in which the host cell is grown. If not secreted, the protein can be isolated from a lysate of the host cells. [0018] In addition to substantially full-length polypeptides used as the source of the selected antigenic amino acid subsequences, biologically active fragments of polypeptides, or analogs thereof, including organic molecules which simulate the interactions of the polypeptides, can be used. Biologically active fragments include any portion of the full-length polypeptide which has a biological function, including ligand binding, and antibody binding. Ligand binding includes binding by nucleic acids, proteins or polypeptides, small biologically active molecules, or large cellular structures. Amino acid sequences identified as antigenic from de novo sequence determination of cDNA reading frames or the isolated protein of interest, or by established sequences from GeneBank and the (PDB), can be most conveniently synthesized by solid phase peptide synthesis using either standard F-Monc or t-Boc methodologies. [0019] This invention also pertains to an isolated polypeptide comprising the antigenic amino acid subsequences of the invention. The encoded proteins or polypeptides of the invention can be partially or substantially purified (e.g., purified to homogeneity), and/or are substantially free of other proteins. According to the invention, the amino acid sequence of the polypeptide can be that of the naturally-occurring polypeptide or can comprise alterations therein. Such alterations include conservative or non-conservative amino acid substitutions, additions and deletions of one or more amino acids; however, such alterations should preserve at least one activity of the encoded protein or polypeptide, i.e., the altered or mutant protein should be an active derivative of the naturally-occurring protein. For example, the mutation(s) can preferably preserve the three dimensional configuration of the binding and/or catalytic site of the native protein, the hydrophilicity and/or flexibility of the polypeptide. The presence or absence of biological activity or activities can be determined by various functional assays as described herein. Moreover, amino acids which are essential for antigenicity or the function of the encoded protein or polypeptide can be identified by methods known in the art. Particularly useful methods include identification of conserved amino acids in the family or subfamily, site-directed mutagenesis and alanine-scanning mutagenesis (for example, Cunningham and Wells, Science 244:1081-1085 (1989)), crystallization and nuclear magnetic resonance. The altered polypeptides produced by these methods can be tested for particular biologic activities, including immunogenicity and antigenicity, as described herein. Continue reading... Full patent description for Anti-chlamydial antibodies and uses thereof Brief Patent Description - Full Patent Description - Patent Application Claims Click on the above for other options relating to this Anti-chlamydial antibodies and uses thereof patent application. ###  How KEYWORD MONITOR works... a FREE service from FreshPatents1. Sign up (takes 30 seconds). 2. Fill in the keywords to be monitored. 3. Each week you receive an email with patent applications related to your keywords. Start now! - Receive info on patent apps like Anti-chlamydial antibodies and uses thereof or other areas of interest. ### Previous Patent Application: Using plasma proteomic pattern for diagnosis, classification, prediction of response to therapy and clinical behavior, stratification of therapy, and monitoring disease in hematologic malignancies Next Patent Application: Methods and compositions involving sortase b Industry Class: Chemistry: molecular biology and microbiology ### FreshPatents.com Support Thank you for viewing the Anti-chlamydial antibodies and uses thereof patent info. IP-related news and info Results in 0.20204 seconds Other interesting Feshpatents.com categories: Novartis , Pfizer , Philips , Polaroid , Procter & Gamble , |
||
